Processes for the Preparation of AZD5363 and Novel Intermediate Used Therein

ABSTRACT

There is provided a process for the preparation of AZD5363, or a salt of AZD5363, comprising: (a) the reaction of 8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione: or a salt thereof, with (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or a salt thereof, in the presence of base; and (b) either isolating AZD5363 or isolating AZD5363 as a salt. 8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione is provided and is prepared by reacting 4-(alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylic acid, or a salt thereof, with a cyclising agent

The present invention relates to chemical processes useful for thepreparation of a certain pharmaceutical compound known as AZD5363. Theinvention also relates to an intermediate compound that has been used aspart of the above-mentioned chemical processes for the improvedpreparation of AZD5363.

The pharmaceutical compound ‘AZD5363’ is alternatively known as:(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide,and its chemical structure is shown below:

International patent application PCT/GB2008/050925 (published asWO2009/047563) mentions AZD5363 as ‘Example 9’ and introduces severalprocesses for its preparation on pages 39-42. The published methods forpreparing AZD5363 were satisfactory for the preparation of relativelysmall quantities. However, clinical trials of AZD5363 have started sincethe filing of WO2009/047563 and in this context, increasing quantitiesof AZD5363 are now required. In our experience, the problems with theexisting methods for preparing AZD5363 include low reaction yields, theformation of impurities and the need for purification steps that are notvery amenable to larger-scale use, the use of chemical reagents andsolvents that are disadvantageous from an environmental and/or safetyand/or cost and/or convenience perspective, relatively long processingtimes for the overall synthesis, relatively large quantities of chemicalwaste per gram of AZD5363 isolated, relatively large cost per gram ofAZD5363 produced, and challenges with ensuring that impurity levels inthe final AZD5363 product are reliably kept to acceptable levels for usein human subjects. Accordingly there is a need for alternative routesfor the preparation of AZD5363 and/or improved processing methods. Oneor more of the above-mentioned needs/problems have been overcome byaspects of the present invention, as described hereinafter.

Although WO2009/047563 discloses several methods for the preparation ofAZD5363, our ongoing internal efforts to deliver AZD5363 for use inclinical trials has been prioritised on the route involving a BOC(t-butoxycarbonyl) deprotection as shown below:

In our hands, on a large scale, this process suffers from poorthroughput and low yields, and operates under high dilution and poorprocess mass intensity. In particular the BOC deprotection step has atedious work-up, involving multiple solvent swaps, poor throughput andthe formation of impurities, despite process development efforts havingbeen focused on these problems. There remained a problem to produceAZD5363 in a way where the above-mentioned problems are overcome orminimized. According to the first aspect of the present invention, asolution to the problem has now been found. This solution does notmerely involve a change of reaction conditions, reagents and/orsolvents, but it also involves the use of an intermediate compound.

Accordingly, in the first aspect of the invention there is provided aprocess for the preparation of AZD5363, or a salt of AZD5363,comprising:

(a) the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione:

or a salt thereof, with (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or asalt thereof, in the presence of base; and(b) either isolating AZD5363 or isolating AZD5363 as a salt.

In one embodiment there is provided a process for the preparation ofAZD5363, comprising the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dioneor a salt thereof, with (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or asalt thereof, in the presence of base.

In a further embodiment there is provided a process for the preparationof AZD5363, comprising the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewith (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, in the presence of base.

In a further embodiment there is provided a process for the preparationof AZD5363, or a salt of AZD5363, comprising:

(a) the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewith (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, in the presence of base;and(b) either isolating AZD5363 or isolating AZD5363 as a salt.

A person skilled in the area of chemistry will recognize that8-(7H-pyrrolo[2,3-d]-pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione,(S)-3-amino-3-(4-chlorophenyl)-propan-1-ol and AZD5363 each possess atleast one nitrogen atom that would be expected to be sufficiently basicto enable the formation of salt forms of the three aforementionedcompounds. Each of these three compounds may be used, generated and/orisolated either in ‘free-base’ form, or in the form of a salt, but theskilled person will recognize that the use of a larger quantity of basemay be appropriate in the above-mentioned process in the case where oneor both of the starting materials are used in the form of a salt.

The above-mentioned process is facilitated by the use of a base. In onerespect, such a base may simply be provided by the presence of one ormore of8-(7H-pyrrolo[2,3-d]-pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionein free-base form, (S)-3-amino-3-(4-chlorophenyl)propan-1-ol infree-base form and/or AZD5363 in free-base form—which may already bepresent in the reaction mixture, although these compounds are relativelyexpensive and the addition of a cheaper base provides a more efficientand/or cost effective process. Organic bases include tertiary aminebases, pyridine and substituted pyridine-based bases such as2,6-lutidine. Examples of cheaper bases that have been used tofacilitate the formation of AZD5363 in the above-mentioned processinclude tertiary amine bases such as triethylamine, N-methylmorpholineand diisopropylethylamine. We have shown that inorganic bases can alsobe used in this process. Examples of inorganic bases include hydroxides(HO), carbonates (CO₃ ²⁻) and bicarbonates (HCO₃ ⁻) of an alkali metalor alkaline earth metal (i.e. the metals found in groups 1 and 2 of theperiodic table), especially the hydroxides, carbonates and bicarbonatesof Li, Na, K, Rb, Cs, Mg, Ca, Sr and Ba.

In one embodiment the base is an organic base or an inorganic base.

In one embodiment the base is selected from a tertiary amine base,pyridine, a substituted pyridine base, and a hydroxide, carbonate orbicarbonate salt of an alkali metal or an alkaline earth metal.

In one embodiment the base is selected from a tertiary amine base,pyridine, a substituted pyridine base, and a hydroxide, carbonate orbicarbonate salt of Li, Na, K, Rb, Cs, Mg, Ca, Sr or Ba.

In one embodiment the base is selected from (C₁₋₆alkyl)₃N,N—(C₁₋₆alkyl)morpholine, N,N—(C₁₋₆alkyl)₂piperazine,N—(C₁₋₆alkyl)piperidine, N—(C₁₋₆alkyl)pyrrolidine, pyridine,2,6-lutidine, 4-(dimethylamino)pyridine and a hydroxide, carbonate orbicarbonate salt of Li, Na, K, Rb, Cs, Mg, Ca, Sr or Ba.

In one embodiment the base is an inorganic base.

In one embodiment the base is an inorganic base selected from ahydroxide, carbonate or bicarbonate salt of an alkali metal or analkaline earth metal.

In one embodiment the base is an inorganic base selected from acarbonate or bicarbonate salt of an alkali metal or an alkaline earthmetal.

In one embodiment the base is an inorganic base selected from ahydroxide, carbonate or bicarbonate salt of Li, Na, K, Rb, Cs, Mg, Ca,Sr or Ba.

In one embodiment the base is an inorganic base selected from acarbonate or bicarbonate salt of Li, Na, K, Rb, Cs, Mg, Ca, Sr or Ba.

In one embodiment the base is an inorganic base selected from acarbonate or bicarbonate salt of Li, Na or K.

In one embodiment the base is an inorganic base selected from abicarbonate salt of Li, Na, or K.

In one embodiment the base is potassium bicarbonate.

In this specification, “C₁₋₆alkyl” means a group containing from 1 to 6carbon atoms where the only other atoms within the group are hydrogenatoms and where the group does not contain any double or triplecarbon-carbon bonds. “C₁₋₆alkyl” includes straight chain alkyl groups,and for C₃₋₆alkyl groups it includes branched chain alkyl groups andalkyl groups that consist of or include a cycloalkyl group. Examples of“C₁₋₆alkyl” include methyl, ethyl, isopropyl, n-butyl, and cyclohexyl.

In this specification “(C₁₋₆alkyl)₃N” means a tertiary amine where eachof the 3 substituents is a C₁₋₆alkyl group, as defined herein. For theavoidance of doubt, the C₁₋₆alkyl groups may each be the same ordifferent when more than one such group is mentioned within a moleculesuch as “(C₁₋₆alkyl)₃N”. Examples of “(C₁₋₆alkyl)₃N” includetriethylamine, diisopropylethylamine and tricyclohexylamine.

Typically, when the8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionehas been isolated as a free base, the reaction with(S)-3-amino-3-(4-chlorophenyl)propan-1-ol may be carried out in thepresence of 1 to 2 mole equivalents of base. In one embodiment, thereaction of the free base of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewith (S)-3-amino-3-(4-chlorophenyl)propan-1-ol may be carried out in thepresence of 1.5 mole equivalents of base. Such reaction may use 1.5 moleequivalent of base, plus or minus 20%. In other embodiments suchreaction may use 1.5 mole equivalent of base, plus or minus 10%.

When the8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionehas been isolated as an acid salt or is prepared in situ using acidicreagents, the reaction with (S)-3-amino-3-(4-chlorophenyl)propan-1-olmay be carried out in the presence of 2 to 3 mole equivalents of base.In one embodiment, the reaction of the acid salt of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewith (S)-3-amino-3-(4-chlorophenyl)propan-1-ol may be carried out in thepresence of 2.5 mole equivalents of base. Such reaction may use 2.5 moleequivalent of base, plus or minus 20%. In other embodiments suchreaction may use 2.5 mole equivalent of base, plus or minus 10%.

It is well known that chemical processes often work more effectivelywhen the reaction components are partially or wholly dissolved into asuitable solvent. In one respect, such solvent used in theabove-mentioned process may simply be provided by the presence of thebase that is already included in the reaction mixture, provided that thebase is a liquid under the relevant reaction conditions. The skilledperson is familiar with a range of solvents that are frequently found tobe suitable for organic chemistry reactions of different types. Typicalsolvents that may be employed for water miscible polar solvents such asdimethylformamide, dimethylacetamide, N-methylpyrollidone, dimethylsulphoxide, sulpholane, tetrahydrofuran, acetonitrile and highernitriles. An example of a suitable solvent is acetonitrile. Mixtures ofsolvents may be used, for example in one embodiment a mixture ofacetonitrile and water is used. A solvent mixture including water may bepreferable when an inorganic base is used as the base, as this may helpensure or encourage dissolution of the inorganic base.

In one embodiment there is provided a process for the preparation ofAZD5363, or a salt of AZD5363, comprising:

(a) the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione:

or a salt thereof, with (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or asalt thereof, in the presence of base and solvent; and(b) either isolating AZD5363 or isolating AZD5363 as a salt.

The processes described herein may provide AZD5363 as a free base, or asa salt if desired. The processes for preparing such a salt are wellknown and may preferably involve the mixture of the amine compound (e.g.AZD5363) with an appropriate quantity (e.g. 1:1 molar ratio) of anorganic or inorganic acid, in a suitable solvent. The resulting salt maysometimes precipitate or crystallize from the solution and in thesecases it may be isolated by filtration. The resulting salt mayalternatively be isolated by evaporation of the solvent.

In our hands, the process for producing AZD5363 from8-(7H-pyrrolo[2,3-d]-pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionecan sometimes proceed less cleanly and/or less completely than desired,depending on the conditions used. It could be speculated that theanhydride functional group within8-(7H-pyrrolo[2,3-d]-pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionemight be susceptible to side reactions, for example reactions withnucleophiles such as hydroxide. Any such reaction could be expected tolead to impurities and lower yields of AZD5363 and might encourage theuse of strictly anhydrous solvents and conditions. However,surprisingly, we have found that the use of water as a co-solvent, incombination with an inorganic base is actually very beneficial forachieving low levels of impurities, improving the reaction rate andhelping to drive the reaction to completion.

Accordingly, in this aspect of the invention there is provided a processfor the preparation of AZD5363, or a salt of AZD5363, comprising:

(a) the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dioneor a salt thereof, with (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or asalt thereof, in the presence of base and solvent; and(b) either isolating AZD5363 or isolating AZD5363 as a salt;wherein the base is one or more inorganic bases and the solvent is amixture of water together with one or more organic solvents wherein thewater comprises between 5% and 50% v/v of the total solvent.

In one embodiment there is provided a process for the preparation ofAZD5363, comprising the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione,or a salt thereof, with (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or asalt thereof, in the presence of base and solvent; wherein the base isone or more inorganic bases and the solvent is a mixture of watertogether with one or more organic solvents wherein the water comprisesbetween 5% and 30% v/v of the total solvent.

In a further embodiment there is provided a process for the preparationof AZD5363, comprising the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione,or a salt thereof, with (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or asalt thereof, in the presence of base and solvent; wherein the base isone or more inorganic bases and the solvent is a mixture of watertogether with one or more organic solvents wherein the water comprisesbetween 7.5% and 22.5% v/v of the total solvent.

In a further embodiment there is provided a process for the preparationof AZD5363, or a salt of AZD5363, comprising:

(a) the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione,or a salt thereof, with (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or asalt thereof, in the presence of base and solvent; and(b) either isolating AZD5363 or isolating AZD5363 as a salt;wherein the base is one or more inorganic bases and the solvent is amixture of water together with one or more organic solvents wherein thewater comprises between 10% and 20% v/v of the total solvent.

8-(7H-Pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionemay be prepared and isolated prior to use in the reaction with(S)-3-amino-3-(4-chlorophenyl)propan-1-ol. Alternatively,8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionemay be prepared in situ and used directly without isolation.

In one embodiment, there is provided a process for the preparation ofAZD5363, or a salt of AZD5363, comprising:

(a) the reaction of compound of4-(alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid, or a salt thereof, with a cyclising agent to give8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione;(b) optionally isolating the8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione;(c) the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewith (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or a salt thereof, inthe presence of base; and(d) either isolating AZD5363 or isolating AZD5363 as a salt.

In one embodiment, there is provided a process for the preparation ofAZD5363, or a salt of AZD5363, comprising:

(a) the reaction of compound of4-(alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid, or a salt thereof, with a cyclising agent to give8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione;(b) reacting the8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewith (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or a salt thereof, inthe presence of base; and then(c) either isolating AZD5363 or isolating AZD5363 as a salt.

In one embodiment, there is provided a process for the preparation ofAZD5363, or a salt of AZD5363, comprising:

(a) the reaction of compound of4-(alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid, or a salt thereof, with a cyclising agent in the presence of afirst solvent to give8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione;(b) optionally isolating the8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione;(c) the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewith (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or a salt thereof, inthe presence of base and second solvent; and(d) either isolating AZD5363 or isolating AZD5363 as a salt.

In one embodiment, there is provided a process for the preparation ofAZD5363, or a salt of AZD5363, comprising:

(a) the reaction of compound of4-(alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid, or a salt thereof, with a cyclising agent in the presence of afirst solvent to give8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione;(b) optionally isolating the8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione;(c) the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewith (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or a salt thereof, inthe presence of base and second solvent; and(d) either isolating AZD5363 or isolating AZD5363 as a salt;wherein the base is one or more inorganic bases and the second solventis a mixture of water together with one or more organic solvents whereinthe water comprises between 5% and 50% v/v of the total solvent.

In a further embodiment, there is provided a process for the preparationof AZD5363, or a salt of AZD5363, comprising:

(a) the reaction of compound of4-(t-butyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid, or a salt thereof, with a cyclising agent in the presence of afirst solvent to give8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione;(b) optionally isolating the8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione;(c) the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewith (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or a salt thereof, inthe presence of base and second solvent; and(d) either isolating AZD5363 or isolating AZD5363 as a salt;wherein the base is one or more inorganic bases and the second solventis a mixture of water together with one or more organic solvents whereinthe water comprises between 10% and 20% v/v of the total solvent.

In one embodiment the4-(alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid is a4-(C₁₋₆-alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid.

In one embodiment the4-(alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid is a4-(C₄-alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid.

In one embodiment the4-(alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid is a4-(t-butyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid.

The first and second solvent may be the same of different and areselected from the solvents listed above.

In one embodiment the first and second solvents are the same.

In one embodiment, where the second solvent is a mixture of more thanone solvent, the first solvent comprises at least one of the solventspresent in the mixture of solvents.

In one embodiment, the first solvent is acetonitrile.

In one embodiment, the second solvent is acetonitrile and water.

In one embodiment, the first solvent is acetonitrile and the secondsolvent is acetonitrile and water.

When the second solvent is acetonitrile and water, in one embodiment thewater comprises between 5% and 50% v/v of the total solvent.

When the second solvent is acetonitrile and water, in another embodimentthe water comprises between 5% and 30% v/v of the total solvent.

When the second solvent is acetonitrile and water, in a furtherembodiment the water comprises between 7.5% and 22.5% v/v of the totalsolvent.

When the second solvent is acetonitrile and water, in a furtherembodiment the water comprises between 10% and 20% v/v of the totalsolvent.

Cyclising agents that may be employed in the reaction of4-(alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid include acid chlorides, acid anhydrides, phosgene equivalents,carbodimides and halotriazines.

Acid chlorides include thionyl chloride, oxayly chloride, pivolylchloride, trichloroacetyl chloride, trifluoroacetyl chloride.

Acid anhydrides include trichloroacetic anhydride, trifluoroaceticanhydride.

Phosgene equivalents include phosgene, diphosgene and triphosgene.

Carbodiimides and halotriazines includeN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide, typically as thehydrochloride salt (EDC.HCl) and 2-chloro-4,6-dimethoxy-1,3,5-triazine(CDMT)

In one embodiment, the cyclising agent is trifluoroacetic anhydride ortrichloroacetyl chloride.

In one embodiment, the cyclising agent is trifluoroacetic anhydride.

In one embodiment, the cyclising agent is trichloroacetyl chloride.

In one embodiment, 1.0 to 1.5 mole equivalents of cyclising agent isused (with respect to the substrate that is being cyclised eg4-(t-butyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid). In a further embodiment 1.3 mole equivalents, plus or minus 10%,cyclising agent is used.

In a further embodiment, there is provided a process for the preparationof AZD5363, or a salt of AZD5363, comprising:

(a) the reaction of compound of4-(t-butyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid with a cyclising agent in the presence of a first solvent to give8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewherein the cyclising agent is trifluoroactetic anhydride and the firstsolvent is acetonitrile;(b) optionally isolating the8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione;and then(c) reacting the8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewith (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, in the presence of baseand second solvent; and(d) either isolating AZD5363 or isolating AZD5363 as a salt;

wherein the base is potassium bicarbonate and the second solvent is amixture of acetonitrile and water wherein the water comprises between10% and 20% v/v of the total solvent.

In a further aspect of the invention there is provided8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione,or a salt thereof.

In a further aspect of the invention there is provided AZD5363 or a saltthereof, obtainable by any of the processes described herein.

In a further aspect of the invention there is provided AZD5363 or a saltthereof, obtained by any of the processes described herein.

In our hands the new methods described to prepare AZD5363 shorten thesynthesis to 3 steps starting from4-(tert-butoxycarbonylamino)piperidine-4-carboxylic acid. The newmethods offer a better isolation procedure for AZD5363. The new methodsoffer the ability to eliminate solvents that are subject to regulatoryrestrictions. The new methods have improved environmental profiles, inparticular the new methods reduce the amount and number of solventsdeployed. The new methods increase overall yields. In addition, thesefactors together lead to an overall cost benefit.

It is understood that AZD5363 prepared by the processes described hereinmay be used to provide formulations such as tablets for use asmedicaments for the treatment of cancer. Suitable formulations andtreatment uses for the medicaments so prepared are described inWO2009/047563.

General Experimental

The invention will now be further explained by reference to thefollowing illustrative examples.

The following abbreviations are used herein or within the followingillustrative examples:—

HPLC High Performance Liquid Chromatography PDA Photodiode ArrayDetector ACN Acetonitrile

DMSO dimethylsulfoxideTFA trifluoroacetic acidThe chemical names were generated by software from ACD labs Version 12.0

Unless stated otherwise, starting materials were commercially available.All solvents and commercial reagents were of laboratory grade and wereused as received.

Stage 1: Water, Acetonitrile, NaOH, 2-MethylTHF. Stage 2: Acetonitrile,Triflouoroacetic Anhydride, Water, KHCO₃, NaOH, Isopropanol Stage 3:Ethanol

EXAMPLE 1. PREPARATION OF4-AMINO-N-[(1S)-1-(4-CHLOROPHENYL)-3-HYDROXY-PROPYL]-1-(7H-PYRROLO[2,3-D]PYRIMIDIN-4-YL)PIPERIDINE-4-CARBOXAMIDE(LEUCHS ANHYDRIDE NON ISOLATED)

To a stirred suspension of4-(t-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (1, 1.0 mol.eq, 46.0 g) in acetonitrile (10.0 rel.vols, 460.0 mL)was slowly added trifluoroacetic anhydride (1.3 mol.eq, 23.26 mL) at 10to 15° C. over a period of 15 to 20 minutes under nitrogen atmosphere.The mixture was stirred for 90 minutes at 25° C. Then potassiumbicarbonate (2.5 mol.eq, 31.86 g) and acetonitrile (3.5 rel.vol, 161.0mL) were added and the reaction mixture stirred for 5-10 mins at 25° C.Then (3S)-3-amino-3-(4-chlorophenyl)propan-1-ol (1.2 mol.eq, 31.47 g)and water (1.5 rel.vols, 69.0 mL) were added and the reaction mixturestirred at 25° C. for 8-10 hrs. Water (5.0 rel.vols, 230.0 mL) wascharged to the reaction mixture and then the mixture was distilled at50-55° C. under reduced pressure, until the residual volume reached 5-6rel.vols, 230.0 mL. Isopropyl alcohol (4.0 rel.vols, 184.0 mL) was addedto the concentrated reaction mass and the pH adjusted to pH˜12.0-12.5using 10% aqueous sodium hydroxide solution (0.7 rel.vols, 32.2 mL) at25° C. The reaction contents were then heated to 55-60° C. and pHre-adjusted to pH˜12.0-12.5 using 10% aqueous sodium hydroxide solution(0.6 rel.vol, 27.6 mL) at 55-60° C. The mixture was stirred for 90 minsand water (10.0 rel.vols, 460.0 mL) was added at 55-60° C. The mixturewas then cooled slowly to 35° C. over a period of 60-90 mins. An aliquotof purified AZD5363 (0.005 mol. Eq, 0.27 g) was added at 35° C. in orderto seed crystallization and the reaction contents were stirred for 30mins. Water (15 rel.vols) was added and stirred for 16-18 hrs at 22-25°C. The mixture was then filtered, and the precipitated solid wasisolated and dried at 60° C. under vacuum to give desired4-amino-N-[(1S)-1-(4-chlorophenyl)-3-hydroxy-propyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperidine-4-carboxamide3 as an white to off white solid with 84.19% yield and Purity: 99% byHPLC area.

Chromatographic Conditions:

Chromatographic separation was achieved on Agilent LC systems equippedwith PDA detectors using Atlantis T3 column and a mixture ofWater:ACN:TFA as an eluent.

¹H NMR-(400.13 MHz, DMSO-d₆) δ: 11.68 (1H, s), 8.48 (1H, d), 8.13(1H,$), 7.37-7.31 (4H, m), 7.16-7.15 (1H, m), 6.57 (1H, m), 4.88 (1H,d), 4.53 (1H, t), 4.41-4.34 (2H, m), 3.59-3.50 (2H, m), 3.40-3.35 (2H,m), 2.17 (2H, s), 2.02-1.80 (4H, m), 1.47-1.39 (2H, m).

EXAMPLE 2: PREPARATION OF4-AMINO-N-[(1S)-1-(4-CHLOROPHENYL)-3-HYDROXY-PROPYL]-1-(7H-PYRROLO[2,3-D]PYRIMIDIN-4-YL)PIPERIDINE-4-CARBOXAMIDE(WITHOUT ISOLATING LEUCHS ANHYDRIDE)

To a stirred suspension of4-(t-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (1, 46.0 g, 127.2 mmole) in acetonitrile (10.0 rel.vols, 460.0 mL)added slowly 2,2,2-trichloroacetyl chloride (1.5 mol.eq, 21.42 mL) at 10to 15° C. over a period of 15 to 20 minutes under nitrogen atmosphere.The reaction contents were stirred for 90 minutes at 25° C. Potassiumbicarbonate (2.5 mol.eq, 31.86 g) and acetonitrile (3.5 rel.vol, 161.0mL) was added to the reaction contents and stirred for 5-10 mins at 25°C., then (3S)-3-amino-3-(4-chlorophenyl)propan-1-ol (1.2 mol.eq, 31.47g) was added and then water (1.5 rel.vols, 69.0 mL) was added and thereaction mixture stirred at 25° C. for 10 hrs. Water (5.0 rel.vols,230.0 mL) was charged to the reaction mixture and the reaction massdistilled at 50-55° C. under reduced pressure, until the residual volumereaches to 5-6 rel.vol. Isopropyl alcohol (4.0 rel.vols, 184.0 mL) wasadded to the concentrated reaction mass and the pH adjusted topH˜12.0-12.5 using 10% w/v sodium hydroxide solution (0.7 rel.vols, 32.2mL) at 25° C. The reaction contents were heated to 55-60° C. and the pHre-adjusted to pH˜12.0-12.5 using 10% w/v sodium hydroxide solution (0.6rel.vol, 27.6 mL) at 55-60° C. The mixture was stirred for 90 mins andwater (10.0 rel.vols, 460.0 mL) was added at 55-60° C. The reactionmixture was cooled slowly to 22-25° C. over the period of 60-90 mins.Water (10.0 rel.vols, 460.0 mL) was added and stirred for 18-20 hrs at22-25° C. The mixture was filtered and the precipitated solid wasisolated and dried at 60° C. under vacuum to give desired4-amino-N-[(1S)-1-(4-chlorophenyl)-3-hydroxy-propyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide3 as an off white solid, 49.0 g (84.19%), Purity (98.24% by HPLC area).

Chromatographic Conditions:

Chromatographic separation was achieved on Agilent LC systems equippedwith PDA detectors using Atlantis T3 column and a mixture ofWater:ACN:TFA as an eluent.

¹H NMR-(400.13 MHz, DMSO-d₆) δ: 11.68 (1H, s), 8.48 (1H, d), 8.13 (1H,s), 7.37-7.31 (4H, m), 7.16-7.15 (1H, m), 6.57 (1H, m), 4.88 (1H, d),4.53 (1H, t), 4.41-4.34 (2H, m), 3.59-3.50 (2H, m), 3.40-3.35 (2H, m),2.17 (2H, s), 2.02-1.80 (4H, m), 1.47-1.39 (2H, m).

EXAMPLE 3. PREPARATION OF4-AMINO-N-[(1S)-1-(4-CHLOROPHENYL)-3-HYDROXY-PROPYL]-1-(7H-PYRROLO[2,3-D]PYRIMIDIN-4-YL)PIPERIDINE-4-CARBOXAMIDE(3) (FROM ISOLATED LEUCHS ANHYDRIDE)

To a stirred suspension of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-2-oxa-4,8-diazaspiro[4.5]-decane-1,3-dionefree base (2, 3.0 g, 10.45 mmole) in acetonitrile (10.0 rel.vols, 30.0mL) was added potassium bicarbonate (2.5 mol.eq, 2.64 g, 26.36 mmole),(3S)-3-amino-3-(4-chlorophenyl)propan-1-ol (1.2 mol.eq, 2.61 g, 12.67mmole), water (1.5 rel.vols, 4.5 mL) and acetonitrile (3.5 rel.vol, 10.5mL). The reaction mixture stirred at 25° C. for 10 hrs. Water (5.0rel.vols, 15.0 mL) was charged to the reaction mixture and the reactionmass distilled at 50-55° C. under reduced pressure, until the residualvolume reaches to 5-6 rel.vols. Isopropyl alcohol (4.0 rel.vols, 12.0mL) was added to the concentrated reaction mass and the pH adjusted topH˜12.0-12.5 using 10% w/v sodium hydroxide solution (0.6 rel.vols, 1.8mL) at 25° C. The reaction mixture was heated to 55-60° C. and the pHre-adjusted to pH˜12.0-12.5 using 10% w/v sodium hydroxide solution (0.6rel.vol, 1.8 mL) at 55-60° C. The mixture was stirred for 90 mins andwater (10.0 rel.vols, 30.0 mL) was added at 55-60° C. The reactionmixture was cooled slowly to 22-25° C. over a period of 60-90 mins.Water (10.0 rel.vols, 30.0 mL) was added and stirred for 18-20 hrs at22-25° C. The mixture was filtered and the precipitated solid isolatedand then dried at 60° C. under vacuum to give desired4-amino-N-[(1S)-1-(4-chlorophenyl)-3-hydroxy-propyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide3 as an off white solid, 3.8 g (80.0%), Purity (99.0% by HPLC area).

Chromatographic Conditions:

Chromatographic separation was achieved on Agilent LC systems equippedwith PDA detectors using Atlantis T3 column and a mixture ofWater:ACN:TFA as an eluent.

¹H NMR-(400.13 MHz, DMSO-d₆) δ: 11.68 (1H, s), 8.48 (1H, d), 8.13(1H,$), 7.37-7.31 (4H, m), 7.16-7.15 (1H, m), 6.57 (1H, m), 4.88 (1H,d), 4.53 (1H, t), 4.41-4.34 (2H, m), 3.59-3.50 (2H, m), 3.40-3.35 (2H,m), 2.17 (2H, s), 2.02-1.80 (4H, m), 1.47-1.39 (2H, m).

Preparation of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-2-oxa-4,8-diazaspiro[4.5]decane-1,3-dione(Leuchs Anhydride Free Base)

To a stirred solution of 2-chloro-4,6-dimethoxy-1,3,5-triazine (1.3mol.eq, 3.04 g) in acetonitrile (10.0 rel.vols, 47.6 mL) was addedN-methylmorpholine (1.3 mol.eq, 1.9 mL) at 22-25° C. The reactioncontents were stirred for 10 mins at 22-25° C.4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (1, 4.76 g, 1.0 mol.eq) was added and the reaction mixture heatedto 45-50° C. The mixture was stirred for 6-8 hrs at 45-50° C. and thencooled to 22-25° C. The mixture was filtered and the precipitated solidisolated to give8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-2-oxa-4,8-diazaspiro[4.5]decane-1,3-dione(2) as a solid; 4.4 g (84.0%), Purity=96.5% by HPLC area.

Chromatographic Conditions:

Chromatographic separation was achieved on Agilent LC systems equippedwith PDA detectors using Atlantis T3 column and a mixture ofWater:ACN:TFA as an eluent.

¹H NMR—(DMSO-d₆) δ: 11.79 (s, 1H), 9.68 (s, 1H), 8.19 (s, 1H), 7.23 (s,1H), 6.63 (s, 1H), 4.37-4.33 (m, 2H), 3.70-3.67 (m, 2H), 2.00-1.92 (m,4H).

¹³C NMR—(DMSO-d₆): 171.7, 154.9, 150.9, 149.5, 149.4, 120.6, 101.2,99.5, 59.7, 40.1, 31.4.

Alternate Preparation of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-2-oxa-4,8-diazaspiro[4.5]-decane-1,3-dione(Leuchs Anhydride Trifluoroacetate Salt) (2)

To a stirred suspension of4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (1, 1.0 mol.eq, 46.0 g) in acetonitrile (10.0 rel.vols, 460.0 mL)was added slowly trifluoroacetic anhydride (1.3 mol.eq, 23.26 mL) at 10to 15° C. over a period of 15 to 20 minutes under nitrogen atmosphere.The reaction mixture was stirred for 90 minutes at 25° C. The mixturewas filtered and the precipitated solid isolated to give8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-2-oxa-4,8-diazaspiro[4.5]decane-1,3-dioneas the trifluoroacetate salt (2) as a solid with 80% isolated yield,Purity=97% by HPLC area.

Chromatographic Conditions:

Chromatographic separation was achieved on Agilent LC systems equippedwith PDA detectors using Atlantis T3 column and a mixture ofWater:ACN:TFA as an eluent.

¹H NMR (DMSO-d₆) δ: 12.52 (s, 1H), 9.73 (s, 1H), 8.35 (s, 1H), 7.39 (s,1H), 6.85 (d, 1H), 4.36 (m, 2H), 3.79 (m, 2H), 2.25-1.93 (m, 4H)

¹³C NMR—(DMSO-d6): 172.6, 159.1, 158.8, 158.5, 158.1, 153.6, 150.6,147.3, 145.6, 123.4, 120.6, 117.7, 114.7, 111.8, 120.6, 102.0, 60.4,42.3, 32.5.

1. A process for the preparation of AZD5363, or a salt of AZD5363,comprising: (a) the reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione:

or a salt thereof, with (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or asalt thereof, in the presence of base; and (b) either isolating AZD5363or isolating AZD5363 as a salt.
 2. A process for the preparation ofAZD5363, or a salt of AZD5363, comprising: (a) the reaction of compoundof4-(alkyloxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid, or a salt thereof, with a cyclising agent to give8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione;(b) reacting the8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dionewith (S)-3-amino-3-(4-chlorophenyl)propan-1-ol, or a salt thereof, inthe presence of base; and then (c) either isolating AZD5363 or isolatingAZD5363 as a salt.
 3. A process according to claim 1 wherein the base isone or more inorganic bases
 4. A process according to claim 3 where thebase is potassium bicarbonate.
 5. A process according to claim 1 whereinthe reaction of8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione,or a salt thereof, with (S)-3-amino-3-(4-chlorophenyl)propan-1-ol is inthe presence of a solvent.
 6. A process according to claim 5 wherein thesolvent is a mixture of acetonitrile and water. 7.8-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-oxa-1,8-diazaspiro[4.5]decane-2,4-dione,or a salt thereof.